title: Assessment of beating parameters in human induced pluripotent stem cells enables quantitative in vitro screening for cardiotoxicity.
abstract: Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of multiple cardiomyocyte physiological parameters enables predictive and mechanistically-interpretable evaluation of cardiotoxicity in a high-throughput format. Human iPSC-derived cardiomyocytes were exposed for 30 min or 24 h to 131 drugs, positive (107) and negative (24) for in vivo cardiotoxicity, in up to 6 concentrations (3 nM to 30 uM) in 384-well plates. Fast kinetic imaging was used to monitor changes in cardiomyocyte function using intracellular Ca(2+) flux readouts synchronous with beating, and cell viability. A number of physiological parameters of cardiomyocyte beating, such as beat rate, peak shape (amplitude, width, raise, decay, etc.) and regularity were collected using automated data analysis. Concentration-response profiles were evaluated using logistic modeling to derive a benchmark concentration (BMC) point-of-departure value, based on one standard deviation departure from the estimated baseline in vehicle (0.3% dimethyl sulfoxide)-treated cells. BMC values were used for cardiotoxicity classification and ranking of compounds. Beat rate and several peak shape parameters were found to be good predictors, while cell viability had poor classification accuracy. In addition, we applied the Toxicological Prioritization Index (ToxPi) approach to integrate and display data across many collected parameters, to derive "cardiosafety" ranking of tested compounds. Multi-parameter screening of beating profiles allows for cardiotoxicity risk assessment and identification of specific patterns defining mechanism-specific effects. These data and analysis methods may be used widely for compound screening and early safety evaluation in drug development.
authors.affiliation: Molecular Devices LLC, Sunnyvale, CA 94089, USA. Electronic address: oksana.sirenko@moldev.com.
authors.forename: Oksana
authors.initials: O
authors.lastname: Sirenko
authors.forename: Evan F
authors.initials: EF
authors.lastname: Cromwell
authors.forename: Carole
authors.initials: C
authors.lastname: Crittenden
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Wignall
authors.forename: Fred A
authors.initials: FA
authors.lastname: Wright
authors.forename: Ivan
authors.initials: I
authors.lastname: Rusyn
mesh_terms: D019540:Area Under Curve; D054715:Cardiotoxins / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions / Q000175:diagnosis*; D006331:Heart Diseases / Q000139:chemically induced / Q000175:diagnosis* / Q000473:pathology; D057166:High-Throughput Screening Assays; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D010641:Phenotype; D018570:Risk Assessment
chemical_list: D054715:Cardiotoxins
keywords: AUC; BMC; Calcium flux; Cardiotoxicity; DMSO; EC(50); Phenotypic screening; Predictive toxicology; area under the curve; benchmark concentration; dimethyl sulfoxide; effective concentration at 50% of the maximum response; hERG; iPSC; induced pluripotent stem cell